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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 王雅筠(Ya-Yun Wang) | |
dc.contributor.author | Hsin-Yi Kuo | en |
dc.contributor.author | 郭馨憶 | zh_TW |
dc.date.accessioned | 2021-06-17T03:46:55Z | - |
dc.date.available | 2019-02-23 | |
dc.date.copyright | 2018-02-23 | |
dc.date.issued | 2018 | |
dc.date.submitted | 2018-01-29 | |
dc.identifier.citation | Abràmoff, M. D., Magalhãe, P. J., and Ram, S. J. (2004). Image processing with ImageJ. Biophotonics International 11, 36-42.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/70162 | - |
dc.description.abstract | 土壤鹽鹼化對農業造成極大的損害,受到影響而減少的糧食產量將是未來人類所要面對的生存難題之一。在鹽分逆境中,茉莉酸訊息傳導路徑會被誘導並且幫助植物調整生存策略以適應新的環境。阿拉伯芥 (Arabidopsis thaliana) 的芥子油苷運輸蛋白 (glucosinolate transporter 1, GTR1/NPF2.10) 屬於硝酸鹽/短胜肽運輸蛋白家族 (nitrate transporter 1/peptide transporter family) 的成員,目前被發現具有運輸硝酸鹽、芥子油苷以及茉莉酸-異亮氨酸複合物 (JA-Ile),GTR1具有兩個可能的磷酸化位點,在過去的發表中,僅提出這兩個磷酸化位點的調控對於芥子油苷的運輸沒有顯著影響,磷酸化對於GTR1的功能具有什麼影響仍是未知。本研究中發現GTR1的表現會受到鹽逆境以及茉莉酸甲酯 (MeJA) 處裡的誘導,但不受到離層酸 (ABA) 的影響,且GTR1在鹽逆境下的表現是受到 JA-Ile 的受體COI1調控的。此外,在鹽逆境及MeJA處理後,gtr1突變株能夠緩解野生型主根生長受到抑制的現象。檢測受鹽逆境所誘導的數個訊息傳導路徑,其中僅茉莉酸訊息傳導路徑在gtr1突變株當中被抑制,而其他路徑在gtr1及野生型當中沒有顯著的差異。這些結果顯示GTR1藉由促進茉莉酸訊息傳導來協助植物應對鹽份逆境。另一方面,本研究也利用非洲爪蟾 (Xenopus) 的卵作為模型探討磷酸化對GTR1運輸功能的影響,試驗結果顯示不同磷酸化程度的GTR1均可以運輸JA-Ile以及硝酸鹽,且Ser635位置的磷酸化可以進一步促進JA-Ile的運輸。有鑑於GTR1能夠運輸多種不同的受質,並且有被磷酸化修飾的可能,GTR1如何參與在鹽份逆境的耐受性調控機制需要更進一步的研究。 | zh_TW |
dc.description.abstract | Salinity stress cause severe loss of crop productivity, and such loss will be a big problem for human beings. Under salinity stress, jasmonate (JA) signaling is activate and helps plants to establish tolerance against unfavorable environment. The Arabidopsis glucosinolate transporter 1 (GTR1/NPF2.10) belongs to nitrate transporter 1/peptide transporter family (NPF), and has been reported to transport nitrate, glucosinolates, and jasmonoyl isoleucine (JA-Ile). In this study, the role of GTR1 on JA signaling under salt stress is investigated. RT-PCR analysis showed that the expression of GTR1 is induced by salt and methyl jasmonate (MeJA) but not by abscisic acid (ABA). In addition, the JA-Ile receptor, COI1, is required for the induction of GTR1 at later stage of salinity stress. In gtr1 mutant, the inhibition of root growth was alleviated under treatment of NaCl or MeJA. The induction of JA responsive genes was inhibited in gtr1 mutant under salt stress while the expression of other salinity responsive genes showed no significant difference between Col-0 and gtr1 mutant. These results suggest that GTR1 participates in the salt stress responses through promoting JA responses. Besides phenotypic analysis, the two putative phosphorylation sites of GTR1 were analyzed for substrate transport activity by Xenopus oocyte system. The results showed that GTR1 and all its phosphorylation mimicries can transport JA-Ile and nitrate, and the phosphorylation on S635 can further promote the JA-Ile transport activity. Due to the multi-substrate property and post-translational modification, the detailed mechanisms of how GTR1 contributes to salt tolerance need further investigation. | en |
dc.description.provenance | Made available in DSpace on 2021-06-17T03:46:55Z (GMT). No. of bitstreams: 1 ntu-107-R04b42017-1.pdf: 1974630 bytes, checksum: 63ffdc94627b96ee63b0d4a799bea0d5 (MD5) Previous issue date: 2018 | en |
dc.description.tableofcontents | 口試委員會審定書 I
致謝 II 摘要 III Abstract IV Contents VI List of figures IX List of supplemental data X List of Appendix XI 1. Introduction 1 1.1. Salinity stress 1 1.2. Jasmonate (JA) and salinity stress 3 1.3. Nitreate transporter 1/peptide transporter family (NPF) 5 1.4. NPF and salt stress 7 1.5. Glucosinolate transporter 1 (GTR1/NPF2.10) 9 2. Specific aims 12 3. Materials and methods 13 3.1. Plant materials and growth conditions 13 3.2. Phenotype analysis 14 3.3. Genome DNA extraction 14 3.4. RNA extraction 16 3.5. Reverse transcription (RT) 17 3.6. Polymerace Chain Reaction (PCR) 18 3.7. Quantitative real-time PCR (qPCR) 19 3.8. PCR/gel purification 19 3.9. Plasmid construction 20 3.10. E. coli. transformation 21 3.11. Plasmid extraction 22 3.12. In vitro transcription 23 3.13. Functional analysis by heterologous expression in Xenopus oocyte 24 3.14. Western blot 26 4. Results 28 4.1. Salinity induces GTR1 expression under the control of COI1. 28 4.2. The growth inhibition of primary root in gtr1 mutant is less sensitive to MeJA and NaCl treatments. 29 4.3. GTR1 participates in JA signaling under salinity stress. 30 4.4. Phosphorylating S635 promotes JA-Ile transport efficiency of GTR1. 32 5. Discussion 33 5.1. GTR1/NPF2.10 is involved in salt-induced inhibition of primary root growth. 33 5.2. Transcriptional regulation of GTR1/NPF2.10 34 5.3. Post-translational regulation of GTR1/NPF2.10 35 5.4. Salt-induced JA responses are different from those induced by MeJA 36 5.5. Roles of GTR1/NPF2.10 in salt responses 37 6. Figures 39 7. Suplemental data 55 8. References 65 9. Appendix 75 9.1. Appendix A: Buffer 75 9.2. Appendix B: Primers and list of constructs 84 | |
dc.language.iso | en | |
dc.title | 阿拉伯芥GTR1/NPF2.10調控鹽逆境誘導茉莉酸訊息傳遞機制之探討 | zh_TW |
dc.title | The roles of GTR1/NPF2.10 in salinity-induced JA signaling in Arabidopsis thaliana | en |
dc.type | Thesis | |
dc.date.schoolyear | 106-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 蔡宜芳(Yi-Fang Tsay),洪傳揚(Chwan-Yang Hong),張英?(Ing-Feng Chang) | |
dc.subject.keyword | 鹽逆境,茉莉酸訊息傳導,NPF,GTR1/NPF2.10,JA-Ile, | zh_TW |
dc.subject.keyword | salt stress,JA signaling,NPF,GTR1/NPF2.10,JA-Ile, | en |
dc.relation.page | 93 | |
dc.identifier.doi | 10.6342/NTU201800212 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2018-01-29 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 植物科學研究所 | zh_TW |
顯示於系所單位: | 植物科學研究所 |
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